Methods of treating protozoal diseases

ABSTRACT

The invention concerns a new pharmaceutical agent for oral or topical administration in the treatment of protozoal diseases, in particular of leishmaniasis, which contains as the active substance one or several compounds having the general formula I  
                 
 
     in which R 1  is a saturated or monounsaturated or polyunsaturated hydrocarbon residue with  12  to  20  C atoms and R 2 , R 3  and R 4  denote independently of one another hydrogen, a C 1 -C 5  alkyl group, a C 3 -C 6  cycloalkyl group or a C 1 -C 5  hydroxyalkyl group whereby two of the residues R 2 , R 3  and R 4  can together form a C 2 -C 5  alkylene group which, if desired, can be substituted with an —O—, —S— or NR 5  group, in which R 5  is hydrogen, a C 1 -C 5  alkyl group, a C 3 -C 6  cycloalkyl group or a C 1 -C 5  hydroxyalkyl group as well as, if desired the usual pharmaceutical auxiliary, diluting, carrier or/and filling substances.

DESCRIPTION

[0001] The present invention concerns a process for the production of apharmaceutical agent for oral or topical administration in the treatmentof protozoal diseases, in particular of leishmaniasis.

[0002] Leishmaniasis is a name for various tropical diseases which arecaused by flagellates of the genus Leishmania and is transmitted byvarious blood-sucking insects. The manifestations of leishmaniasis maybe visceral (kala-azar), mucocutaneous (american leishmaniasis) orcutaneous (Aleppo boil or diffuse cutaneous leishmaniasis). Theincubation period is weeks or months. A very high mortality rate isobserved in untreated cases, in particular with kala-azar and americanleishmaniasis.

[0003] The therapeutic agents used today for the treatment ofleishmaniasis are pentavalent antimony compounds (e.g. sodiumstibogluconate) and aromatic diamidines. A disadvantage of these drugsis, however, that they cause severe side-effects such as nausea andvomitting due to their high toxicity. Moreover there are alreadyLeishmania strains which are resistant to antimony.

[0004] Croft et al (Biochem. Pharmacol. 36 (1987), p. 2633-2636)describe experiments in which the effectiveness of alkyl phosphocholinesagainst Leishmania donovani was investigated. The effect of alkylphosphocholines was tested in comparison with the effect of the standardtherapeutic preparation, sodium stibogluconate, (Pentostam) whenadministered subcutaneously. In this process it was found that alkylphosphocholines, in particular C₂₂ alkyl phosphocholines are activeagainst Leishmania. It was, however, also established that the alkylphosphocholines, in particular hexadecyl-phosphocholine, were highlytoxic for the experimental animals, in particular for macrophages intherapeutically effective doses so that they do not represent a realalternative when administered subcutaneously to the known therapy withsodium stibogluconate.

[0005] Thus the object of the present invention was to provide apharmaceutical agent for protozoal diseases, in particular forleishmaniasis, in which the disadvantages of the state of the art, inparticular with regard to the severe side-effects, are at leastpartially eliminated.

[0006] The object according to the present invention is achieved by aprocess for the production of a pharmaceutical agent for oral or topicaladministration in the treatment of protozoal diseases, in particular ofleishmaniasis, which contains as the active substance one or severalcompounds having the general formula I

[0007] in which R¹ is a saturated or monounsaturated or polyunsaturatedhydrocarbon residue with 12 to 20 C atoms and R², R³ and R⁴ denoteindependently of one another hydrogen, a C₁-C₅ alkyl group, a C₃-C₆cycloalkyl group or a C₁-C₅ hydroxyalkyl group whereby two of theresidues R², R³ and R⁴ can together form a C₂-C₅ alkylene group which,if desired, can be substituted with an —O—, —S— or NR⁵ group, in whichR⁵ is hydrogen, a C₁-C₅ alkyl group, a C₃-C₆ cycloalkyl group or a C₁-C₅hydroxyalkyl group as well as, if desired the usual pharmaceuticalauxiliary, diluting, carrier or/and filling substances.

[0008] In the general formula I, R¹ can be branched or straight-chained.R¹ is preferably a straight-chained hydrocarbon residue with 16 to 20 Catoms, in particular a hexadecyl, octadecyl, oleyl, elaidyl, eicosyl oreicosenyl-cis-(ω-9) residue. R¹ is particularly preferably a hexadecylor octadecyl residue.

[0009] Examples of suitable residues R², R³ and R⁴ in the formula I arefor instance methyl, ethyl, propyl, butyl and pentyl residues,cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl residues,hydroxymethyl, hydroxyethyl and hydroxypropyl residues. Two of theresidues R², R³ and R⁴ can for example form a pyrrolidine, a piperidineor a morpholine group. At least one of the residues R², R³ and R⁴ ispreferably different from hydrogen, it is particularly preferred thatall 3 residues are different from hydrogen.

[0010] Examples of preferred residues R², R³ and R⁴ are methyl, ethyl,hydroxyethyl and C₃-C₆ cycloalkyl residues. If one of R², R³ and R⁴ is acycloalkyl residue, then the other two residues are preferably methylresidues. It is particularly preferred that the residues R², R³ and R⁴are independently of each other methyl or ethyl residues. It is mostpreferred when R², R³ and R⁴ are methyl residues so that alkylphosphocholines represent a particularly preferred class of compoundswhich is suitable for the production of an agent against protozoaldiseases, in particular against leishmaniasis.

[0011] It was surprisingly found that compounds having the generalformula I when administered orally or topically show no measurableside-effects and a very much higher activity than sodium stibogluconate.In any case the therapeutic agents according to the present inventionconstitute the first forms of oral therapy for leishmaniasis diseases,and they are considerably more effective than Pentostam, a standardtherapeutic preparation used worldwide in the liver and in particularalso in the spleen.

[0012] In a preferred embodiment of the present invention the oral ortopical therapeutic preparation additionally contains one or severalalkyl glycerols having the general formula II

[0013] in which one of the residues R⁶ and R⁷ denotes an alkyl groupwith 2 to 12 C atoms and the other residue denotes a hydrogen atom. Analkyl glycerol mixture is preferably used which contains nonyl or octylglycerol, hexyl or pentyl glycerol and propyl or ethyl glycerol as wellas, if desired, water.

[0014] The pharmaceutical agent according to the present inventioncontains in one dosage unit preferably 5 to 2000 mg, particularlypreferably 10 to 500 mg of one or several compounds having the generalformula I. For topical administration the pharmaceutical agent accordingto the present invention preferably contains 5 to 200 mg of one orseveral compounds having the general formula I per ml of an alkylglycerol having the formula II or of a corresponding alkyl glycerolmixture.

[0015] For oral administration the pharmaceutical agent according to thepresent invention is preferably formulated as a drinking solution with adaily dosage between 1 and 10 mg/kg of one or several compounds havingthe general formula I.

[0016] The production of an oral pharmaceutical agent according to thepresent invention can on the other hand also be carried out by mixing orhomogenizing one or several compounds having the general formula I withthe usual physiologically tolerated filling, carrier, dilution or/andauxiliary substances at temperatures between 20 and 120° C. and, ifdesired in order to prepare formulations which contain 10 to 800 mg ofcompounds having the general formula I in one dosage unit, the mixturethus obtained is poured into hollow cells of an appropriate size orfilled into capsules of an appropriate size or granulated and thenpressed into tablets, if desired, with addition of further commonauxiliary substances. The active substance can for example be mixed withone or several of the following auxiliary substances: starch, cellulose,lactose, formalin-casein, modified starch, magnesium stearate, calciumhydrogenphosphate, highly-dispersed silicic acid, talcum andphenoxyethanol. The mixture obtained is granulated, if desired, with anaqueous solution containing for example gelatin, starch, polyvinylpyrrolidone, vinylpyrrolidon-vinyl acetate copolymerisate or/andpolyoxyethylene sobitanmonooleate, as constituent and the granulate ishomogenized, if desired, with one or several of the aforementionedauxiliary substances. Subsequently this mixture can be pressed intotablets or filled into capsules whereby the tablets or capsules eachcontain 10 to 800 mg of active substance in one dosage unit.

[0017] In a particularly preferred embodiment the active substance issuspended with soybean lecithin as well as, if desired, 0.1 to 0.5 partsby weight phenoxyethanol (in relation to one part by weight of theactive substance) at temperatures between 33 and 37° C. in melted resinfat and homogenized and subsequently the mixture is poured into hollowcells whereby one dosage unit contains 10 to 800 mg of the activesubstance.

[0018] Moreover the active substance can be homogenized at a temperaturebetween 50 and 120° C., if desired in the presence of one or severalemulsifiers or/and 0.1 - 0.5 parts by weight phenoxyethanol (in relationto 1 part by weight of the active substance) with at least one of thefollowing substances: paraffin, vaseline, aliphatic alcohol with 12 to25 C atoms, sorbitanmonopalmitate, aliphatic monocarboxylic acid with 15to 20 C atoms, polyoxyethylenepolyol fatty acid ester. If desired, themixture obtained can be emulsified with addition of a multivalent lower(preferably C₂-C₃) aliphatic alcohol (e.g. ethylene glycol, diethyleneglycol, propylene glycol, dipropylene glycol and in particularglycerol).

[0019] On the other hand, if desired, the active substance can bedissolved at temperatures between 30 and 100° C. in the presence of0.1 - 0.5 parts by weight phenoxyethanol (in relation to one part byweight of the active substance) as well as, if desired, in the presenceof an emulsifier and if desired the solution thus obtained is filled upwith sufficient water or vegetable oil so that the final solutioncontains 0.1 to 5 % by weight of the active substance.

[0020] The active substance can also be mixed together with an alkylglycerol having the general formula II or with a mixture of such alkylglycerols as well as, if desired, water in which 1 to 30 parts by weightalkyl glycerol having the general formula II or of a corresponding alkylmixture and if desired 1 to 30 parts by weight water, each in relationto 1 part by weight active substance according to general formula I, areused.

[0021] The present invention also concerns the use of one or severalcompounds having the general formula I as the active substance for anoral or a topical agent for treating protozoal diseases, in particularleishmaniasis. In this connection the agent can additionally contain oneor several alkyl glycerols having the general formula II, in particularfor topical applications.

[0022] Finally the invention also concerns a process for the treatmentof protozoal diseases, in particular of leishmaniasis, which ischaracterized in that a pharmaceutical agent produced according to thepresent invention is administered topically or orally.

[0023] The production of compounds having the general formula I isdescribed in detail in the examples for hexadecyl-phosphocholine.Further methods for the production of compounds having the generalformula I are described for example in DE-A 27 52 125, DE-A 36 41 379,DE-A 36 41 491, DE-A 40 13 632, DE-A 36 41 377, the literature cited inthese or in earlier patent applications or patent specifications of thesame applicant. Reference is expressly made to this literature for thepresent patent application.

[0024] The pharmaceutical agents according to the present invention arepreferably used for the treatment of leishmaniasis. Other protozoaldiseases which can be treated by the agent according to the presentinvention are for instance malaria, trypanosomiasis, toxoplasmosis,babesiosis, amoebic dysentery and lambliasis. The agents according tothe present invention are in particular suitable for those diseases inwhich the pathogen is present in organs such as the liver, spleen orkidney.

[0025] It is intended to elucidate the invention further by thefollowing examples.

EXAMPLE 1 Production of Hexadecylphosphocholine H₂O a)Hexadecylphosphoethanolamine (Phosphorylation, Ring Closure and RingOpening)

[0026] Hexadecanol (1 mole, 243 g) and triethylamine (1.8 mole, 180 g)are dissolved in 1.5 l THF (tetrahydrofuran) and added dropwise to asolution of phosphoroxychloride (1.2 mole, 184 g) in 120 ml THF whichwas stirred vigorously in such a way that the temperature in thereaction vessel (three-neck, 5 l, with dropping funnel, thermometer andstirrer) does not exceed 10° C. In order to accelerate the process, thereaction vessel is cooled with an ice-salt mixture. The reaction iscompleted immediatly after the dropwise addition (detected by TLC inether: Rf values of 0.8 for the initial product, of 0.0 for the reactionproduct after hydrolysis with water).

[0027] The ice-bath is removed and a solution of ethanolamine (1.5 mole,92 g) and triethylamine (1.8 mole, 180 g) in 1 l dioxan are added whilestirring vigorously in such a way that the temperature in the reactionvessel increases to 65 to 70° C. Then the ring formation is completed(detected by TLC in ether: Rf values of 0.2). Precipitated triethylaminehydrochloride is removed by filtration while still warm and 1.5 l 2Nformic acid is added to the filtrate at 40 to 50° C. After 15 minutesthe ring opening is completed (detected by TLC in ether: Rf values 0.0;TLC in chloroform/methanol/acetic acid/water 100:60:20:5 by volume: Rfvalue 0.8). It is cooled to −20° C. and the precipitate which is mainlycomposed of pure hexadecylphosphoethanolamine is filtered off. If slightimpurities are present a subsequent chromatographic purification iscarried out. Microanalysis (MW 365.50): calc. (%) C 59.15 H 11.03 N 3.83P 8.48 found (%) 59.01 10.95 3.79 8.31

[0028] Methylation of hexadecylphosphoethanolamine

[0029] The crystals obtained according to example 1 are taken up in 1.2l 2-propanol and 0.4 l dichloromethane without further purification.Potassium carbonate (4 mole, 560 g) is added to the suspension of thecrystals in 1 l water while stirring vigorously. Dimethyl sulfate (4mole, 500 g) is added dropwise to the two-phase reaction mixture whilestirring in such a way that the temperature does not exceed 40° C. Thereaction is completed 60 minutes after the dropwise addition (detectedby TLC in chloroform/methanol/25% ammonia 50:50:5 by volume: Rf value0.3). After phase separation at 20° C., the upper phase contains theproduct. The solvent is removed in a rotary evaporator under a vacuumand the viscous residue is chromatographed on silica gel (Merck productNo. 7733, silica gel 60, particle size 0.2 to 0.5 mm).

Chromatography

[0030] Chloroform/methanol/25% ammonia (200/15/1 by volume) is added to2 kg silica gel and filled into a chromatography column. The viscous oilis dissolved in 800 ml of the above solvent mixture and the crudeproduct is applied to the column (insoluble components are previouslyremoved by filtration). It is eluted with mobile solvents of increasingpolarity until the impurities are washed out. The product is finallyeluted with chloroform/methanol/25% ammonia (50/50/5 by volume). Thecombined eluates are rotary evaporated and the remaining water isremoved with toluol. The residue is taken up in 600 ml dichloromethaneand 4 l acetone is added. The crystals which separate out at −20° C. arewashed with cold acetone, then with pentane and dried in a vacuum. Theyield of pure hexadecylphospocholine is 250 g (ca. 70% in relation tohexadecylglycerol). Microanalysis (MW 407.58): calc. (%): C 59.27 H11.37 N 3.29 P 7.28 found (%): 58.98 11.31 3.21 7.11

[0031] Production of pharmaceutical formulations

Example for a Solution

[0032] 25 g 1-n-propyloxy-2,3-propanediol, 12.5 g1-n-hexyloxy-2,3-propanediol, 12.5 g 1-n-nonyloxy-2,3-propanediol, 44 gwater and 1 g phenoxyethanol are mixed and 5 g hexadecylphosphocholineis dissolved in this mixture. The solution is freed of visible particlesby filtration over suitable filters.

[0033] 1 g solution contains 50 mg hexadecylphosphocholine.

Example for a an Ointment

[0034] 5 g hexadecylphosphocholine is suspended in 35 g viscousparaffin, 30 g emulsified cetylstearyl alcohol and 30 g white vaselineare added and melted. This melt is first stirred until it has cooleddown. A homogeneous distribution of active substance is achieved byprocessing the cooled melt by means of a suitable homogenizer (e.g.three-roll mill).

[0035] 1 g of the hydrophilic ointment contains 50 mghexadecylphosphocholine.

Example for an Emulsion

[0036] 11.83 g 1-n-propyloxy-2,3-propanediol, 5.91 g1-n-hexaloxy-2,3-propanediol, 5.91 g 1-n-nonyloxy-2,3-propanediol, 20.35g water and 1.0 g phenoxyethanol are mixed and 5 ghexadecylphosphocholine is dissolved in this mixture. 30 g whitevaseline, 15 g cetylalcohol and 5 g sorbitan monopalmitate are melted ona water-bath, heated to 70° C. and the solution of the active substance,which was also heated to 70° C., is emulsified in the fat phase with theaid of a high-speed dispersing apparatus. The cream is subsequentlycooled down to 30° C. while stirring. 1 g water-in-oil cream contains 50mg hexadecylphosphocholine.

Example for Capsules

[0037] 1.25 kg hexadecylphospocholine is dissolved in 5 kg chloroformand 1.25 kg aerosil is suspended in this solution. The solvent issubsequently removed in a vacuum. The dry mass is passed through a 1 mmsieve and dried once again in a vacuum at 30° C. in order to remove anylast remains of solvent. This granulate is filled in a known way to 500mg into gelatin hard capsules with a size of 00 using a suitable capsulemachine. One capsule contains 250 mg hexadecylphosphocholine.

Examples of Further Active Substances EXAMPLE 2

[0038] Octadecylphosphocholine

[0039] C₂₃H₅₀NO₄P MW 435.630

EXAMPLE 3

[0040] Oleylphosphocholine

[0041] C₂₃H₄₈NO₄P MW 433.614

EXAMPLE 4

[0042] Elaidylphosphocholine

[0043] C₂₃H₄₈NO₄P MW 433.614

EXAMPLE 5

[0044] Hexadecylphospho-(N.N-dimethyl-N-ethyl)ethanolamine MW 421.603

EXAMPLE 6

[0045] Octadecylphospho-(N.N-dimethyl-N-ethyl)ethanolamine MW 449.657

EXAMPLE 7

[0046] Oleylphospho-(N.N-dimethyl-N-ethyl)ethanolamine MW 447.641

EXAMPLE 8

[0047] Elaidylphospho-(N.N-dimethyl-N-ethyl)ethanolamine MW 447.641

EXAMPLE 9

[0048] Hexadecylphospho-(N-cyclopropyl-N.N-dimethyl)-ethanolamine

[0049] C₂₃H₄₈NO₄P MW 443.614

EXAMPLE 10

[0050] Hexadecylphospho-(N-cyclobutyl-N.N-dimethyl)-ethanolamine

[0051] C₂₄H₅₀NO₄P MW 447.641

EXAMPLE 11

[0052] Hexadecylphospho-(N-cyclopentyl-N.N-dimethyl)-ethanolamine

[0053] C₂₅H₅₂NO₄P MW 461.668

EXAMPLE 12

[0054] Hexadecylphospho-(N.N-dimethyl-N-hydroxyethyl)-ethanolamine

[0055] C₂₂H₄₈NO₅P MW 437.602

EXAMPLE 13

[0056] Hexadecylphospho-(N-methyl)-pyrrolidino-ethyl ester

[0057] C₂₃H₄₈NO₄P MW 433.614

EXAMPLE 14

[0058] Octadecylphospho-(N.N-diethyl-N-methyl)-ethanolamine

[0059] C₂₅H₅₄NO₄P MW 463.684

EXAMPLE 15

[0060] Octadecylphospho-(N-cyclopropyl-N.N-dimethyl)-ethanolamine

[0061] C₂₅H₅₂NO₄P MW 461.668

EXAMPLE 16

[0062] Octadecylphospho-(N-cyclobutyl-N.N-dimethyl)-ethanolamine

[0063] C₂₆H₅₄NO₄P MW 475.695

EXAMPLE 17

[0064] Octadecylphospho-(N-cyclopentyl-N.N-dimethyl)-ethanolamine

[0065] C₂₇H₅₆NO₄P MW 489.722

EXAMPLE 18

[0066] Octadecylphospho-(N.N-dimethyl-N-hydroxyethyl)-ethanolamine

[0067] C₂₄H₅₂NO₄P MW 465.656

EXAMPLE 19

[0068] Octadecylphospho-(N-methyl)-pyrrolidino-ethyl ester

[0069] C₂₅H₅₂NO₄P MW 461.668

EXAMPLE 20

[0070] Oleylphospho-(N.N-diethyl-N-methyl)-ethanolamine

[0071] C₂₅H₅₂NO₄P MW 461.668

EXAMPLE 21

[0072] Oleylphospho-(N-cyclopropyl-N.N-dimethyl)-ethanolamine

[0073] C₂₅H₅₀NO₄P MW 459.652

EXAMPLE 22

[0074] Oleylphospho-(N-cyclopentyl-N.N-dimethyl)-ethanolamine

[0075] C₂₇H₅₄NO₄P MW 487.706

EXAMPLE 23

[0076] Oleylphospho-(n.N-dimethyl-N-hydroxyethyl)-ethanolamine

[0077] C₂₄H₅₀NO₅P MW 463.640

EXAMPLE 24

[0078] Oleylphospho-(N-methyl)-pyrrolidino-ethyl ester

[0079] C₂₅H₅₀NO₄P MW 459.652

EXAMPLE 25

[0080] Elaidylphospho-(N-cyclopropyl-N.N-dimethyl)-ethanolamine

[0081] C₂₅H₅₀NO₄P MW 459.652

EXAMPLE 26

[0082] Elaidylphospho-(N.N-dimethyl-N-hydroxyethyl)-ethanolamine

[0083] C₂₄H₅₀NO₅P MW 463.640

EXAMPLE 27

[0084] Elaidylphospho-(N-methyl)-pyrrolidino-ethyl ester

[0085] C₂₅H₅₀NO₄P MW 459.652

EXAMPLE 28

[0086] Eicosylphosphocholine

[0087] C₂₅H₅₄NO₄P MW 463.684

EXAMPLE 29

[0088] Eicosylphospho-(N-ethyl-N.N-dimethyl)-ethanolamine

[0089] C₂₆H₅₆NO₄P MW 477.711

EXAMPLE 30

[0090] Eicosylphospho-(N.N-diethyl-N-methyl)-ethanolamine

[0091] C₂₇H₅₄NO₄P MW 491.738

EXAMPLE 31

[0092] Eicosylphospho-(N-cyclopropyl-N.N-dimethyl)-ethanolamine

[0093] C₂₇H₅₆NO₄P MW 489.722

EXAMPLE 32

[0094] Eicosylphospho-(N.N-dimethyl-N-hydroxyethyl)-ethanolamine

[0095] C₂₆H₅₆NO₅P MW 493.710

EXAMPLE 33

[0096] Eicosylphospho-(N.N-dihydroxyethyl-N-methyl)-ethanolamine

[0097] C₂₇H₅₈NO₆P MW 523.736

EXAMPLE 34

[0098] Eicosylphospho-(N-methyl)-pyrrolidino-ethyl ester

[0099] C₂₇H₅₆NO₄P MW 489.722

EXAMPLE 35

[0100] Eicosenyl-cis-(ω-9)-phosphocholine

[0101] C₂₅H₅₂NO₄P MW 461.668

EXAMPLE 36

[0102] Eicosenyl-cis-(ω-9)-phospho-(N-ethyl-N.N-dimethyl)-ethanolamine

[0103] C₂₆H₅₄NO₄P MW 475.695

EXAMPLE 37

[0104]Eicosenyl-cis-(ω-9)-phospho-(N-cyclopropyl-N.N-dimethyl)-ethanolamine

[0105] C₂₇H₅₄NO₄P MW 487.706

EXAMPLE 38

[0106]Eicosenyl-cis-(ω-9)-phospho-(N.N-dimethyl-N-hydroxyethyl)-ethanolamine

[0107] C₂₆H₅₄NO₅P MW 491.694

EXAMPLE 39

[0108] Effect of various leishmaniasis drugs on the presence ofpathogens in the liver of experimental animals (rats) infected with L.donovani.

[0109] The effect of the phospholipids according to the presentinvention (hexadecylphosphocholine, octadecylphospho-choline andoleylphospho-(N.N-dimethyl-N-ethyl)-ethanolamine) was compared with thestandard therapeutic agent Pentostam used worldwide and the ether lipidEt₁₈OCH₃ (1-octadecyl-2-methyl-rac-glycero-3-phosphocholine).

[0110] The compounds according to the present invention and Et₁₈OCH₃were administered orally while Pentostam was administered intravenously.It was found that the alkyl phosphocholines with a C₁₈ and C₁₆ chainwere 32-times more effective than the standard therapeutic agentPentostam while olely-phospho-(N.N-dimethyl-N-ethyl)ethanolamine has acomparable effect to Pentostam.

[0111] The results of this experiment are shown in Table 1. The numberof pathogens per liver was determined by microscopic analysis. TABLE 1Number of leishmania pathogens in the liver after a 3 week period oftherapy Relative effectiveness (pathogens per liver after Pathogens perPentostam therapy divided liver (in by pathogens/liver after millions)addition of test substance) Control 536.9 — 1-octadecyl-2-methyl- 424.70.007 rac-glycero-3-phospho- choline¹ Oleylphospho-(N.N- 4.7 0.7dimethyl-N-ethyl)- ethanolamine¹ Pentostam² 3.2 1.0 Hexadecylphospho-0.1 32.0 choline¹ Octadecylphospho- 0.1 32.0 choline¹

[0112] In some additional experiments parasites were no longermicroscopically detectable in the liver and in the spleen after singleoral dose of 200 mg hexadecylphosphocholine.

EXAMPLE 40

[0113] Effect of different leishmaniasis drugs on the presence ofpathogens in the spleen

[0114] The experimental procedure was as described in example 39. Theresults of these experiments are shown in Table II. TABLE II Number ofleishmania pathogens in the spleen after a 3 week period of therapyRelative effectiveness (pathogens per spleen after Pathogens perPentostam therapy divided spleen (in by pathogens/spleen after millions)addition of test substance) Control 24.3 — 1-octadecyl-2-methyl 6.5 0.9rac-glycero-3-phospho- choline Oleylphospho-(N.N- 0.03 210.0dimethyl-N-ethyl)- ethanolamine¹ Pentostam 6.3 1.0 Hexadecylphospho-0.01 630.0 choline Octadecyiphospho- 0.01 630.0 choline

[0115] The weak effect of Pentostam in the spleen is surprising. Thealkylphosphocholines are in this case >600-fold more effective than thestandard therapy.

[0116] In several additional experiments parasites were no longermicroscopically detectable after a single oral dose of 200 mghexadecylphosphocholine.

[0117] Table III shows the concentration of hexadecyl-phosphocholine(C₁₆—O—PC), octadecylphosphocholine (C₁₈—O—PC) and1-octadecyl-2-methyl-rac-glycero-3-phosphocholine (Et₁₈OCH₃) in theorgans of the rat after oral administration of 50 μmol/day for 5 days, apause of 2 days then for a further 4 days at 50 μmol/day. TABLE IIIAmount of substance (nmol/g fresh tissue) Organ C₁₆-O-PC C₁₈-O-PCEt₁₈OCH₃ Serum 130 47 5 Liver 272 298 36 Spleen 410 424 43 Kidney 853406 57

[0118] It is surprising how well hexadecylphosphocholine andoctadecylphosphocholine accumulate in the spleen. It is especially inthe spleen that the standard therapeutic agent Pentostam has anextremely low effectiveness. In comparison to this Et₁₈OCH₃ is onlypresent in the spleen and in the liver in extremely low concentrations.

1. Process for the production of a pharmaceutical agent for oral ortopical administration in the treatment of protozoal diseases, inparticular of leishmaniasis, which contains as the active substance oneor several compounds having the general formula I

in which R¹ is a saturated or monounsaturated or polyunsaturatedhydrocarbon residue with 12 to 20 C atoms and R², R³ and R⁴ denoteindependently of one another hydrogen, a C₁-C₅ alkyl group, a C₃-C₆cycloalkyl group or a C₁-C₅ hydroxyalkyl group whereby two of theresidues R², R³ and R⁴ can together form a C₂-C₅ alkylene group which,if desired, can be substituted with an —O—, —S— or NR⁵ group, in whichR⁵ is hydrogen, a C₁-C₅ alkyl group, a C₃-C₆ cycloalkyl group or a C₁-C₅hydroxyalkyl group as well as, if desired the usual pharmaceuticalauxiliary, diluting, carrier or/and filling substances.
 2. Process asclaimed in claim 1 , wherein R¹ is straight-chained and has 16 to 20 Catoms.
 8. Process as claimed in claim 7 , wherein the pharmaceuticalagent contains an alkyl glycerol mixture of nonyl or octyl glycerol,hexyl or pentyl glycerol and propyl or ethyl glycerol as well as, ifdesired, water.
 9. Process as claimed in one of the previous claims,wherein the pharmaceutical agent contains 5 to 2000 mg, in particular 10to 500 mg, of one or several compounds having the general formula I inone dosage unit.
 10. Process as claimed in one of the previous claims,wherein it contains as a topical pharmaceutical agent 5 - 200 mg of oneor several compounds having the general formula I per ml of an alkylglycerol having the formula II or of a corresponding alkyl glycerolmixture.
 11. Process as claimed in one of the claims 1 to 10 , whereinit is formulated as a drinking solution having a daily dose between 1and 10 mg/kg of one or several compounds having the general formula I.12. Process as claimed in one of the claims 1 to 10 , wherein one orseveral compounds having the general formula I are mixed or homogenizedwith the usual physiologically tolerated filling, carrier, dilutionor/and auxiliary substances at temperatures between 20 and 120° C. and,if desired, in order to prepare formulations which contain 10 to 800 mgof compounds having the general formula I in one dosage unit, themixture thus obtained is poured into hollow cells of an appropriate sizeor filled into capsules of an appropriate size or granulated and thenpressed into tablets, if desired, with addition of further commonauxiliary substances.
 13. Process as claimed in claim 12 , wherein oneor several compounds having the general formula I are mixed with one orseveral of the following auxiliary substances: starch, cellulose,lactose, formalin-casein, modified starch, magnesium stearate, calciumhydrogenphosphate, highly-dispersed silicic acid, talcum andphenoxyethanol, the mixture obtained is granulated if desired with anaqueous solution containing the constituents gelatin, starch, polyvinylpyrrolidone, vinylpyrrolidone-vinyl acetate copolymerisate or/andpolyoxyethylene sobitanmonooleate, the granulate is homogenized, ifdesired, with one or several of the aforementioned auxiliary substancesand this mixture is pressed into tablets or filled into capsules wherebythe tablets or capsules each contain 10 to 800 mg of compounds havingthe general formula I in one dosage unit.
 14. Process as claimed in oneof the claims 12 or 13, wherein one or several compounds having thegeneral formula I are suspended with soybean lecithin as well as ifdesired, 0.1 to 0.5 parts by weight phenoxyethanol (in relation to onepart by weight of compounds having the general formula I) attemperatures between 33 - 37° C. in melted resin fat and homogenized andsubsequently the mixture is poured into hollow cells whereby the dosageunit contains 10 to 800 mg of compounds having the formula I. 15.Process as claimed in one of the claims 12 or 13, wherein, one orseveral compounds having the general formula I are homogenized at atemperature between 50 and 120° C., if desired, in the presence of oneor several emulsifiers or/and 0.1 - 0.5 parts by weight phenoxyethanol(in relation to 1 part by weight of compounds having the formula 1) withat least one of the following substances: paraffin, vaseline, aliphaticalcohols with 12 to 25 C atoms, sorbitanmonopalmitate, aliphaticmonocarboxylic acid with 15 to 20 C atoms, polyoxyethylene polyol fattyacid ester, and the homogenized mixture is emulsified, if desired, withaddition of a multivalent lower aliphatic alcohol.
 16. Process asclaimed in one of the claims 12 to 15 , wherein one or several compoundshaving the general formula I are dissolved at temperatures between 30 -100° C., if desired, in the presence of 0.1 - 0.5 parts by weightphenoxyethanol (in relation to one part by weight of compounds havingthe formula I) as well as, if desired, in the presence of an emulsifierand if desired, the solution thus obtained is filled up with sufficientwater or vegetable oil so that the final solution contains 0.1 - 5percent by weight of compounds having the formula I.
 17. Process asclaimed in one of the claims 7 to 16, wherein an alkyl glycerol isadditionally used in the production having the formula II

in which one of the residues R⁶ and R⁷ denotes an alkyl group with 2 to12 C atoms and the other residue denotes a hydrogen atom, or a mixtureof such alkyl glycerols as well as, if desired, water is used whereby1 - 30 parts by weight of an alkyl glycerol having the formula II or ofa corresponding alkyl glycerol mixture and if desired 1 - 30 parts byweight of water, each in relation to 1 part by weight of compoundshaving the formula I, are used.
 18. Use of one or several compoundshaving the general formula I defined in claim 1 for the treatment ofprotozoal diseases, in particular of leishmaniasis.
 19. Use as claimedin claim 18, wherein the agent additionally contains one or severalalkyl glycerols having the general formula II defined in claim 7 . 20.Process for the treatment of protozoal diseases, in particular ofleishmaniasis, wherein a pharmaceutical agent as claimed in one of theclaims 1 to 17 is administered topically or orally.